AGILENT ASSAYMAP SOLUTION FOR AFFINITY PURIFICATIONQuantitative, reproducible affinity purification that integrates with other sample prep applications

AssayMAP Affinity Purification Solution

AssayMAP Affinity Purification Solution, from Agilent Technologies, is quantitative front-end solution to protein sample preparation workflows. It is based on the powerful combination of miniaturized, Protein A and G packed bed chromatography cartridges, the state-of-the art Bravo Liquid Handling Platform and a simple applications-based user interface.

With the AssayMAP Affinity Purification solution, 8-96 samples may be processed in parallel using either direct binding of a target antibody or by immobilizing an antibody to capture its antigen. It may be used as a standalone application or integrated with other AssayMAP applications such as In-Solution Digestion and Peptide Cleanup to automate and streamline the entire workflow for LC/MS analysis. AssayMAP workflows dramatically improve reproducibility while decreasing hands on time.

Highlights

• Walkaway automation

• Excellent analytical precision – 5% CVs

• Quantitative recovery from 1 to 100 µg

• Powerful enrichment – recover sample in 10 μL of eluate

• Easy integration with other AssayMAP workflows

Figure 1. Affinity Purification workflows

Product description Part numberAssayMAP Protein A Cartridge rack G5496-60000 1 rack of 96 cartridges

AssayMAP Protein G Cartridge rack G5496-60008 1 rack of 96 cartridges

Ordering Information

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Total Cartridge Elution Volume, µL

PG-W PA-W

www.agilent.com/lifesciences/AssayMAP

This item is intended for Research Use Only. Not for use in diagnostic procedures. This

information is subject to change without notice.

© Agilent Technologies, Inc. 2014 Published in the USA, May 26, 2014

5991-4604EN

hIgG loaded on cartridge (µg)

% recovery %CV – 24 samples

100 91.0 1.1

50 92.3 1.8

25 93.5 1.8

12.5 95.4 1.3

• 12.5, 25, 50, and 100 μg of hIgG were each spiked into 24 replicates of CCS, bound to 96 PG-W cartridges, and eluted with 5% acetic acid into a UV plate for A280 quantitation.

• CVs were below 2% for each sample load mass

• Recovery was greater than 90% for each load mass

A. Varying masses of hIgG in cell culture supernatant were purified using AssayMAP Protein A (PA-W) and Protein G (PG-W) cartridges. The mass of hIgG in the eluate was quantified and plotted against the mass of the load. The identity line is shown.

B. 96 replicates of 100 µg hIgG were purified using PA-W cartridges. The data demonstrates the reproducibility of recovery.

Quantitative binding and elution with minimal elution volumes. CA B

Excellent recovery and analytical precision across a wide mass range.

C. hIgG was bound to PA-W and PG-W cartridges and then subject to isocratic elution with 5% acetic acid. 2 µL fractions were collected and quantified by absorbance. The cumulative elution volume is plotted against the cumulative percent recovery of the load mass. Greater than 95% of the load was recovered in 10 µL of eluate.

Samples can be eluted directly into the appropriate solution for downstream processing.

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%CV = 1.26

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human IgG load, µg

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